Vasopressin Regulates Extracellular Vesicle Uptake by Kidney Collecting Duct Cells
Vasopressin Regulates Extracellular Vesicle Uptake by Kidney Collecting Duct Cells
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DOI：
10.1681/asn.2015050568
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发表时间：
2016-11-01
影响因子：
13.6
通讯作者：
Dear, James W.
中科院分区：
文献类型：
--
作者：
Oosthuyzen, Wilna;Scullion, Kathleen M.;Dear, James W.
细胞外囊泡（ECV）促进沿着肾单位的细胞间通讯，有可能改变受体细胞的功能。然而，尚不清楚这是否是类似于其他信号系统的受监管过程。我们研究了 ECV 转移的潜在激素调节，并报道去氨加压素（一种加压素类似物）刺激肾集合管细胞系 (mCCD(C11)) 和原代细胞摄取荧光负载的 ECV。将 mCCD(C11) 细胞暴露于从过表达 microRNA-503 的细胞中分离出的 ECV 中，会导致 microRNA-503 靶基因的表达下调，但仅限于去氨加压素存在的情况下。从机制上讲，ECV 进入 mCCD(C11) 细胞需要产生 CAMP，通过抑制动力来减少 CAMP 的产生，并且对来自肾小管细胞的 ECV 具有选择性。在体内，我们测量了在给予加压素 V2 受体拮抗剂托伐普坦之前和之后全身递送给小鼠的荧光负载 ECV 的尿液排泄和组织摄取。在对照组治疗的小鼠中，我们在尿液中回收了 2.5% 的 ECV；托伐普坦使恢复率提高了五倍，并减少了肾组织中的 ECV 沉积。此外，在患有中枢性尿崩症的患者中，去氨加压素减少了来自肾小球和近端肾小管细胞的 ECV 的排泄。这些数据与体内加压素调节的 ECV 摄取一致。我们的结论是 ECV 的吸收是一个特定的、受监管的过程。从生理学上来说，ECV是一种新的细胞间通讯机制；在治疗上，ECV可能是一种载体，通过RNA疗法可以靶向特定细胞来治疗肾脏疾病。
Extracellular vesicles (ECVs) facilitate intercellular communication along the nephron, with the potential to change the function of the recipient cell. However, it is not known whether this is a regulated process analogous to other signaling systems. We investigated the potential hormonal regulation of ECV transfer and report that desmopressin, a vasopressin analogue, stimulated the uptake of fluorescently loaded ECVs into a kidney collecting duct cell line (mCCD(C11)) and into primary cells. Exposure of mCCD(C11) cells to ECVs isolated from cells overexpressing microRNA-503 led to downregulated expression of microRNA-503 target genes, but only in the presence of desmopressin. Mechanistically, ECV entry into mCCD(C11) cells required CAMP production, was reduced by inhibiting dynamin, and was selective for ECVs from kidney tubular cells. In vivo, we measured the urinary excretion and tissue uptake of fluorescently loaded ECVs delivered systemically to mice before and after administration of the vasopressin V2 receptor antagonist tolvaptan. In control-treated mice, we recovered 2.5% of administered ECVs in the urine; tolvaptan increased recovery five-fold and reduced ECV deposition in kidney tissue. Furthermore, in a patient with central diabetes insipidus, desmopressin reduced the excretion of ECVs derived from glomerular and proximal tubular cells. These data are consistent with vasopressin-regulated uptake of ECVs in vivo. We conclude that ECV uptake is a specific and regulated process. Physiologically, ECVs are a new mechanism of intercellular communication; therapeutically, ECVs may be a vehicle by which RNA therapy could be targeted to specific cells for the treatment of kidney disease.